An integrated bioanalytical method development and validation approach: case studies

Abstract

We proposed an integrated bioanalytical method development and validation approach: (1) method screening based on analyte's physicochemical properties and metabolism information to determine the most appropriate extraction/analysis conditions; (2) preliminary stability evaluation using both quality control and incurred samples to establish sample collection, storage and processing conditions; (3) mock validation to examine method accuracy and precision and incurred sample reproducibility; and (4) method validation to confirm the results obtained during method development. This integrated approach was applied to the determination of compound I in rat plasma and compound II in rat and dog plasma. The effectiveness of the approach was demonstrated by the superior quality of three method validations: (1) a zero run failure rate; (2) >93% of quality control results within 10% of nominal values; and (3) 99% incurred sample within 9.2% of the original values. In addition, rat and dog plasma methods for compound II were successfully applied to analyze more than 900 plasma samples obtained from Investigational New Drug (IND) toxicology studies in rats and dogs with near perfect results: (1) a zero run failure rate; (2) excellent accuracy and precision for standards and quality controls; and (3) 98% incurred samples within 15% of the original values.

Validation of a high‐performance liquid chromatography method for determining lysophosphatidylcholine content in bovine pulmonary surfactant medication

“…Subsequently, detection was carried out using a simple and cost‐effective HPLC setup that is widely available in most chromatography laboratories. Finally, owing to the importance of method validation (Beg et al, 2016; Brown, 2024; Chen et al, 2024; Chintala, 2024; Dar et al, 2022; Davis et al, 2024; Heudi & Winter, 2024; Hussien, 2014; Rajadhyaksha & Londhe, 2024; Sharma et al, 2012; Xue et al, 2012), the method underwent comprehensive validation for eight parameters. Importantly, the execution of this analytical method does not depend on any state‐of‐the‐art equipment.…”